Neuro Oncol. 2013 Jul 21. [Epub ahead of print]
Erlotinib resistance in EGFR-amplified glioblastoma cells is associated with upregulation of EGFRvIII and PI3Kp110d
Schulte A, Liffers K, Kathagen A, Riethdorf S, Zapf S, Merlo A, Kolbe K, Westphal M, Lamszus K.
Department of Neurosurgery, University Hospital Hamburg-Eppendorf, Hamburg, Germany (A.S., K.L., A.K., S.Z., K.K., M.W., K.L.); Institute for Tumor Biology, University Hospital Hamburg-Eppendorf, Hamburg, Germany (S.R.); Laboratory of Molecular Neuro-Oncology, Department of Clinical and Biological Sciences, University of Basel, Basel, Switzerland (A.M.).
BackgroundThe treatment efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors like erlotinib has not met expectations for glioblastoma therapy, even for EGFR-overexpressing tumors. We determined possible mechanisms of therapy resistance using the unique BS153 glioblastoma cell line, which has retained amplification of the egfr gene and expression of EGFR variant (v)III.MethodsFunctional effects of erlotinib, gefitinib, and cetuximab on BS153 proliferation, migration, and EGFR-dependent signal transduction were systematically compared in vitro. The tumor-initiating capacity of parental and treatment-resistant BS153 was studied in Naval Medical Research Institute/Foxn1nu mice. Potential mediators of resistance were knocked down using small interfering (si)RNA.ResultsErlotinib and gefitinib inhibited proliferation and migration of BS153 in a dose-dependent manner, whereas cetuximab had no effect. BS153 developed resistance to erlotinib (BS153resE) but not to gefitinib. Resistance was associated with strong upregulation of EGFRvIII and subsequent activation of the phosphatidylinositol-3-OH kinase (PI3K) pathway in BS153resE and an increased expression of the regulatory 110-kDa delta subunit of PI3K (p110d). Knockdown of EGFRvIII in BS153resE largely restored sensitivity to erlotinib. Targeting PI3K pharmacologically caused a significant decrease in cell viability, and specifically targeting p110d by siRNA partially restored erlotinib sensitivity in BS153resE. In vivo, BS153 formed highly invasive tumors with an unusual growth pattern, displaying numerous satellites distant from the initial injection site. Erlotinib resistance led to delayed onset of tumor growth as well as prolonged overall survival of mice without changing tumor morphology.ConclusionsEGFRvIII can mediate resistance to erlotinib in EGFR-amplified glioblastoma via an increase in PI3Kp110d. Interfering with PI3Kp110d can restore sensitivity toward the tyrosine kinase inhibitor. |
